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ORIGINAL ARTICLE
Year : 2014  |  Volume : 7  |  Issue : 2  |  Page : 129-137

Use of nested PCR-RFLP for genotyping of Cryptosporidium parasites isolated from calves and children suffering from diarrhea


Department of Parasitology, Faculty of Medicine, Menoufia University, Menoufia, Egypt

Correspondence Address:
Gehan S Sadek
10th Ahmed Zaky Street, El Noza El Gedeeda, 11769, Cairo
Egypt
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/1687-7942.149568

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Background The vast majority of human cases of cryptosporidiosis worldwide are caused by two species: Cryptosporidium hominis (C. parvum type 1), which causes infection in humans only, and Cryptosporidium parvum (C. parvum type 2), which causes infections in humans and animals. In Egypt, calves carrying the zoonotic C. parvum represent the largest farm animal source of infection for humans. Information on the source of Cryptosporidium spp. contamination is necessary for effective evaluation and selection of management practices for reducing the risk for cryptosporidiosis. Objective The aim of the study was to genotypically characterize Cryptosporidium spp. in a sample of isolates from calves and children suffering from diarrhea. Materials and methods One hundred stool samples were collected from diarrheic calves housed at the Tropical Diseases Clinic, Faculty of Veterinary Medicine, Cairo University. A total of 110 stool samples were also collected from diarrheic children attending the Gastroenteritis Unit, Abo El Reesh Pediatric Hospital, Cairo University. Each stool sample of children or calves was examined microscopically after staining with modified Ziehl-Neelsen stain for the diagnosis of Cryptosporidium spp. Positive samples were then subjected to nested PCR-restriction-fragment length polymorphism (PCR-RFLP) targeting Cryptosporidium oocyst wall protein (COWP) gene for determination of Cryptosporidium genotypes. Results Screening by modified Ziehl-Neelsen staining detected Cryptosporidium in stools of 40% of diarrheic calves. Nested PCR-RFLP analysis showed that all positive samples were related to C. parvum genotype 2 (C. parvum). In diarrheic children, screening diagnosed 12/110 (10.9%) positive cases; 9/12 (75%) of them were confirmed positive by nested PCR. RFLP analysis showed that 8/9 (88.9%) samples were C. parvum genotype 1 (C. hominis), whereas one sample was not digested. Conclusion Genotype 2 C. parvum is relatively highly prevalent in the sample of calves examined compared with genotype 1 in the sample of children, indicating that transmission of cryptosporidiosis among this sample of children is anthroponotic and not zoonotic. It is advised to include PCR-RFLP technique in routine clinical diagnosis and epidemiological investigations.


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